Submitted to: Entomologia Experimentalis et Applicata
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/19/2003
Publication Date: 5/1/2004
Citation: Gregory, P.G., Rinderer, T.E. 2004. Non-destructive dna sampling in honey bee queens, apis mellifera. Entomologia Experimentalis et Applicata. Vol. 111:173-177. Interpretive Summary: DNA Technology (gene identification or marker assisted breeding) has great potential for improving honey bee stocks. However, marking must be done very early in the life of a honey bee queen, preferably before she becomes an adult, before marker assisted breeding is of any practical value. Larval and pupal skins deposited during molting were evaluated as sources of DNA to evaluate markers early in a queen¿s life. These procedures proved to be successful. Honey bee breeders are now positioned to take full advantage of the revolution in DNA technology in their breeding efforts.
Technical Abstract: Extracting DNA from Apis mellifera L. queen honey bees can be accomplished without causing harm to the queen. Here we present a simple Chelex¿ extraction method of obtaining DNA from the last larval instar and pupal exuvia, and demonstrate that this DNA can be used to amplify microsatellite regions. DNA was also extracted from wing clippings, another non-invasive source of DNA. DNA from larval castings, pupal exuvia and wing clippings gave genotypes consistent with DNA extracted from the tarsus. DNA analysis of the larval exuvia provides a potential method of genotyping queens before they become adult, a valuable tool for developing a marker-assisted breeding program for honey bees. Utilizing the larval skins provides a way to obtain queen genotypes far enough in advance of adult emergence to allow only selected queen cells to be introduced to colonies, facilitating unrestricted breeder queen production. Thus, early genotyping allows the selection of queens before any major resources are expended. DNA from wing clippings, or larval and pupal castings can be used for genotyping queens that will be instrumentally inseminated in scientific experiments. Also, exceptional queens already established in hives but not yet genotyped can be analyzed from wing clippings.